老年2型糖尿病病人心血管病死亡相关外周血全基因组DNA甲基化的研究

Analysis of genomic DNA methylation of peripheral blood in elderly patients with type 2 diabetes mellitus suffering form cardiovascular disease-induced death

目的利用全基因组DNA甲基化芯片技术,分析老年T2DM心血管病死亡病人和非死亡病人外周血全基因组DNA甲基化基因的差异,寻找与老年T2DM病人心血管病死亡相关的异常甲基化位点。方法选取南京医科大学附属老年医院糖尿病分阶段达标管理(China Staged Diabetes Targeting Management,SDTM)项目截至2017年12月底结局事件为心血管病死亡的12例T2DM病人为病例组,采用倾向性评分法进行对照的匹配,卡钳值设置为0.02选取同期仍存活的12例T2DM病人作为对照组。使用Illumina Methylation BeadChip芯片技术检测外周血全基因组DNA甲基化水平。采用DAVID数据库对差异具有统计学意义的甲基化位点基因进行基因本体(GO)和京都基因与基因组百科全书(KEGG)信号通路分析。结果共发现差异甲基化位点303个。GO功能分析显示,差异甲基化位点基因参与的生物学过程主要有(1)神经系统发育、细胞黏附分子、血糖稳态和树突发育正调控等生物过程;(2)质膜的组成成分、树突和细胞质核周区细胞成分;(3)钙离子结合和序列特异性DNA结合分子功能。KEGG信号通路分析结果显示,差异甲基化基因主要参与细胞内环磷酸腺苷(cAMP)信号通路和炎症介质对瞬时受体电位离子(TRP)通道的调节通路。结论DNA甲基化修饰可能与T2DM病人的心血管病死亡结局相关。

Objective A genome-wide DNA methylation microarray technique was used to explore the aberrantly methylated sites in the elderly patients with type 2 diabetes mellitus(T2DM)suffering from cardiovascular disease-induced death.Methods The present study was conducted using the data from the Staged Diabetes Targeting Management(SDTM)study.The propensity score method was used to match the control,and the calipers value was set to 0.02.Twelve elderly patients with T2DM who died of cardiovascular disease until October 2017 were selected as case group.Twelve elderly patients with T2DM who survived in the same period were selected as the control group.Whole genome methylation level in periphral blood was detected by Illumina Methylation BeadChip.Gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)signaling pathway analysis were performed using the DAVID database.Results A total of 303 CpG sites with statistically significant differences were found.GO function analysis showed that methylation variation positions were involved in such biological process as(1)Nervous system development,cell adhesion,glucose homeostasis and positive regulation of dendrite development;(2)Cellular component such as integral component of plasma membrane,dendrite and perinuclear region of cytoplasm;(3)Molecular function such as calcium ion binding and sequence-specific DNA binding.KEGG signaling pathway analysis found that methylation variation positions were involved in cyclic adenosine monophosphate(cAMP)signaling pathway and inflammatory mediator regulation of transient receptor potential(TRP)channels.Conclusions DNA methylation modification may be involved in the pathogenesis of the elderly patients with T2DM who die of cardiovascular disease.